Modulation of mechanosensitive genes during embryonic aortic arch development.

BACKGROUND: Early embryonic aortic arches (AA) are a dynamic vascular structures that are in the process of shaping into the great arteries of cardiovascular system. Previously, a time-lapsed mechanosensitive gene expression map was established for AA subject to altered mechanical loads in the avian embryo. To validate this map, we investigated effects on vascular microstructure and material properties following the perturbation of key genes using an in-house microvascular gene knockdown system. RESULTS: All siRNA vectors show a decrease in the expression intensity of desired genes with no significant differences between vectors. In TGFß3 knockdowns, we found a reduction in expression intensities of TGFß3 (≤76%) and its downstream targets such as ELN (≤99.6%), Fbn1 (≤60%), COL1 (≤52%) and COL3 (≤86%) and an increase of diameter in the left AA (23%). MMP2 knockdown also reduced expression levels in MMP2 (≤30%) and a 6-fold increase in its downstream target COL3 with a decrease in stiffness of the AA wall and an increase in the diameter of the AA (55%). These in vivo measurements were confirmed using immunohistochemistry, western blotting and a computational growth model of the vascular extracellular matrix (ECM). CONCLUSIONS: Localized spatial genetic modification of the aortic arch region governs the vascular phenotype and ECM composition of the embryo and can be integrated with mechanically-induced congenital heart disease models.

Mechanical characterization and torsional buckling of pediatric cardiovascular materials.

In complex cardiovascular surgical reconstructions, conduit materials that avoid possible large-scale structural deformations should be considered. A fundamental mode of mechanical complication is torsional buckling which occurs at the anastomosis site due to the mechanical instability, leading surgical conduit/patch surface deformation. The objective of this study is to investigate the torsional buckling behavior of commonly used materials and to develop a practical method for estimating the critical buckling rotation angle under physiological intramural vessel pressures. For this task, mechanical tests of four clinically approved materials, expanded polytetrafluoroethylene (ePTFE), Dacron, porcine and bovine pericardia, commonly used in pediatric cardiovascular surgeries, are conducted (n = 6). Torsional buckling initiation tests with n = 4 for the baseline case (L = 7.5 cm) and n = 3 for the validation of ePTFE (L = 15 cm) and Dacron (L = 15 cm and L = 25 cm) for each are also conducted at low venous pressures. A practical predictive formulation for the buckling potential is proposed using experimental observations and available theory. The relationship between the critical buckling rotation angle and the lumen pressure is determined by balancing the circumferential component of the compressive principal stress with the shear stress generated by the modified critical buckling torque, where the modified critical buckling torque depends linearly on the lumen pressure. While the proposed technique successfully predicted the critical rotation angle values lying within two standard deviations of the mean in the baseline case for all four materials at all lumen pressures, it could reliably predict the critical buckling rotation angles for ePTFE and Dacron samples of length 15 cm with maximum relative errors of 31% and 38%, respectively, in the validation phase. However, the validation of the performance of the technique demonstrated lower accuracy for Dacron samples of length 25 cm at higher pressure levels of 12 mmHg and 15 mmHg. Applicable to all surgical materials, this formulation enables surgeons to assess the torsional buckling potential of vascular conduits noninvasively. Bovine pericardium has been found to exhibit the highest stability, while Dacron (the lowest) and porcine pericardium have been identified as the least stable with the (unitless) torsional buckling resistance constants, 43,800, 12,300 and 14,000, respectively. There was no significant difference between ePTFE and Dacron, and between porcine and bovine pericardia. However, both porcine and bovine pericardia were found to be statistically different from ePTFE and Dacron individually (p < 0.0001). ePTFE exhibited highly nonlinear behavior across the entire strain range [0, 0.1] (or 10% elongation). The significant differences among the surgical materials reported here require special care in conduit construction and anastomosis design.

A Portable Smartphone-based Platform with an Offline Image-processing Tool for the Rapid Paper-based Colorimetric Detection of Glucose in Artificial Saliva.

In this study, a microfluidic paper-based analytical device (µPAD) was integrated with a smartphone app capable of offline (without internet access) image processing and analysis for the rapid colorimetric detection of glucose. A self-inking stamp was used to form hydrophobic channels on a piece of paper-towel due to its superior water absorption efficiency. As demonstrated, the developed sensor was employed for the colorimetric detection of glucose in artificial saliva in the linear scope of 0 - 1 mM with a calculated detection limit of 29.65 µM. The experimental results show that the quantitative analysis of glucose with the proposed smartphone platform could be completed in less than one minute. The app developed for the smartphone platform is capable of extracting the color-changing area with an embedded image processing tool which could address the problem of color uniformity in the detection zones of µPAD. The integrated platform has great potential to be used for non-invasive measurements of glucose in body fluids, like tears, sweat and saliva.

Guiding neural extensions of PC12 cells on carbon nanotube tracks dielectrophoretically formed in poly(ethylene glycol) dimethacrylate.

The PC12 cell line has been widely used as an in vitro model for studying neuronal differentiation and identifying the factors affecting the process. It has the ability to differentiate in the presence of nerve growth factor (NGF), resulting in neural extensions called dendrites and axons. In this study, first the impact of randomly distributed multi-walled carbon nanotubes (MWCNTs) in poly(ethylene glycol) dimethacrylate (PEGDMA) on PC12 cell differentiation was investigated in terms of neurite length, number of neurite per cell and differentiation marker gene expression profile. Then, dielectrophoretically aligned MWCNTs in PEGDMA was used to guide and support the neuronal differentiation of PC12 cells in the presence of NGF. The method is expected to be useful in revealing the nanotopographical role in fundamental studies and understanding of nanotopographical effects for biomedical applications on nerve regeneration.